Enterovirus detection in stool specimen: relevance for poliovirus and enterovirus surveillance.

S Haddad-Boubaker, A Ben Yahia, D Rezig, W Farès, H Touzi, H Triki
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Abstract

Detection of enterovirus genome by PCR in clinical samples is now extensively used for the diagnostic of enterovirus infections given its rapidity and high sensitivity. In contrast, its use in surveillance programs targeting specific enterovirus serotypes remains less frequent. The most sensitive protocols are those amplifying in the 5'untranslated region (5'UTR). However the possibility to use sequence analysis of the 5'UTR amplicons for serotype identification is not yet well established. In this report, stool samples from polio suspected cases and their healthy contacts were tested. The results of direct detection of enterovirus genome by PCR and serotype identification based on sequence analysis of the PCR products in the 5'UTR were compared to those of standard cell-culture-based protocols. Standard protocols detected enterovirus isolates in 7.4% of cases while 9.8% of samples were positive by PCR. Serotype identification based on sequence analysis of amplicons showed concordant results with serotypes determined on virus isolates by seroneutralisation or sequencing in the VP1 gene in 39% of cases only. These results confirm that the use of PCR amplification from stool samples improves the sensitivity of enterovirus detection but do not recommend the use of sequence analysis of the 5'UTR PCR product to determine enterovirus serotype.

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粪便标本肠病毒检测:与脊髓灰质炎病毒和肠病毒监测的相关性。
PCR检测肠道病毒基因组具有快速、灵敏等优点,目前已广泛应用于临床标本肠病毒感染的诊断。相比之下,它在针对特定肠道病毒血清型的监测项目中的应用仍然较少。最敏感的协议是那些扩增在5'非翻译区(5'UTR)。然而,使用5'UTR扩增子的序列分析进行血清型鉴定的可能性尚未很好地建立。在本报告中,对脊髓灰质炎疑似病例及其健康接触者的粪便样本进行了检测。将PCR直接检测肠道病毒基因组和基于5'UTR PCR产物序列分析的血清型鉴定结果与基于标准细胞培养方案的结果进行比较。标准方案在7.4%的病例中检出肠道病毒分离株,9.8%的样本经PCR检测呈阳性。基于扩增子序列分析的血清型鉴定结果显示,只有39%的病例与通过血清中和或VP1基因测序确定的病毒分离株血清型一致。这些结果证实,使用粪便样本的PCR扩增提高了肠道病毒检测的敏感性,但不建议使用5'UTR PCR产物的序列分析来确定肠道病毒的血清型。
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